Acute toxicity test in DDY mice of ethanolic extract of irradiated Clinacanthus nutans Lindau leaves

Snake grass (Clinacanthus nutans Lindau) (Dandang gendis in Indonesian) belongs to the Acanthaceae and has been known by people for curing the various diseases, such as diabetes, dysentery, Aedes aegypti larvicide, herpes simplex antiviral antioxidants, antitumor, etc. Preservation of herbal medicine simplicia for a certain period can cause microbial contamination on simplicia. Gamma irradiation technique at the dose of 5 – 10 kGy is one of the post-harvest handlings of dried herbal medicine simplicia, so that the shelf life of the commodity can be extended and it not easily damaged by insects and microbial spoilage. However, there is not much information about the acute oral toxicity of irradiated of dried C. nutans leaves. The study aimed to determine the acute toxicity potent of gamma irradiated at 7.5 kGy and un-irradiated C. nutans leaves using male and female DDY strain mice based on the procedure of the Organization for Economic Cooperation and Development (OECD) and the National Agency of Drug and Food Control (BPOM RI). DDY strain mice aged 8 weeks, weight of 24.1 – 39.4g as test animals were fasted for 8 h before and 4 h after being given the test samples. The sample concentration used were 50, 300, 2000, and 5000 mg/kg BW. The observation was done during 14 d, including toxic symptoms, death of test animals, body weight (average daily gain) and manifestations of toxic effects. At the end of the test, the mice were sacrificed and the vital organs were observed for pathological and histological abnormalities. The results showed that the oral administration of the ethanolic extract of C. nutans leaves (from the irradiated and un-irradiated sample as control) at the test dose limit of 2000 mg/kg BW did not show any toxic symptoms or death of the mice. Moreover, gross pathological observation of vital organs did not show reveal any changes at each dose, while the microscopic examination found thickening of the pulmonary alveolus walls, and mild congestion in the spleen and kidney organs. Based on the classification category of the acute toxic class by BPOM RI No. 7 of 2014 and OECD, the findings of the toxicity study were classified as “practically non-toxic". It meant that gamma irradiation up to the dose of 7.5 kGy to extend the shelf life of the G. nutans simplicia did not cause the simplicia to become acutely toxic. However, further study on chemical components and their biological activities are required.